JP2010540572A - ムユヨの花、トゲバンレイシの葉、及びウコンの根の抽出物を含む、肝炎を治療するための組成物 - Google Patents
ムユヨの花、トゲバンレイシの葉、及びウコンの根の抽出物を含む、肝炎を治療するための組成物 Download PDFInfo
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Abstract
【選択図】 なし
Description
本発明のハーブ組成物の抽出物は、それらのin vitroでの抗酸化作用を決定するために、初代ラット肝細胞培養を用いてテストした。一般的に、テストは、肝細胞の単離が実行されるまでthe Fe de Valencia Hospital Research Centerで飼育されていた、CRIFFA(Santa Perpetua de La Mogoda、バルセロナ)によって提供された雄ラット(Sprague Dawley)を用いて実行した。動物の取扱い及び屠殺は、国家規制、欧州連合の規定609/86、及びUS国立衛生研究所(NIH)によって刊行された実験動物の取扱い及び使用に従って実行した。
細胞生存率は、淡黄色物質であるジメチル−テトラゾリウム(MTT)の取り込み及び青色不溶性代謝産物であるホルマザンへの還元を測定することによって決定した。この細胞還元反応は、ミトコンドリア脱水素酵素活性を測定しており、この活性は細胞内小器官の完全性の程度に依存しており、従って培養中の生存細胞の数の明確な指標となる。この場合において、MTTは黄色を示すテトラゾリウム塩物質として使用され、ミトコンドリア脱水素酵素コハク酸活性によって青色不溶性代謝産物(ホルマザン)を産生し、これは、405〜630nmの範囲の吸光度を測定するELISAリーダーを用いて吸光度を測定することによって定量化され得る。
フリーラジカルの産生を定量化するために、96ウェルプレートに培養されたラット肝細胞の初代培養は、その無極性及びその非イオン性構造のおかげで細胞膜を通じてよく拡散し、酸素が存在すると蛍光を発する蛍光剤である5−クロロメチル−2’,7’−ジクロロヒドロフルオレセイン(DCFH−DA)と共に、以下の濃度:4、20、100及び500μg/mlでの本発明のハーブ組成物とインキュベートした(Lautraite S,Bigot−Lasserre D,Bars R and Carmichael N.Optimisation of cell−based assays for medium throughput screening of oxidative stress.Toxicol in vitro 17:207−220(2003)、これはこの参照によって本明細書に組込まれるものである)。インキュベーション期間が完了したら、細胞を本発明のハーブ組成物と共にt−ブチルヒドロペルオキシド(t−BOOH)に曝露し、その後すぐ蛍光発光を485nm(刺激)及び527nm(発光)で判読した(t0)。最終的に前記細胞は37℃でインキュベートし、蛍光は30分間隔で2時間判読した。ケルセチン(抗酸化作用を有することが知られているフラボノイド)で処理した細胞はテストのポジティブコントロールとして使用した(Boots A.W.,Bast A.and Haenen G.R.No role of DT−diaphorase(NqO1)in the protection against oxidized quercetin.FEBS Lett 579:677−682、これはこの参照によって本明細書に組込まれるものである)。定量化は、コントロール(ハーブ組成物の非存在下で酸化的刺激によって誘導された細胞)に対するフリーラジカルの%で示した。フリーラジカルを定量化するために、ハーブ組成物は0.5〜1%の範囲でエタノール−水溶液中に第一に溶解した。
脂質過酸化及びグルタチオン還元を測定するために、単離ラット肝細胞を80,000生細胞/cm2の密度で24ウェルプレートに培養した。1時間培養の安定化期間の後、細胞は4、20、100及び500μg/mlのハーブ組成物と共に24時間プレインキュベートした。プレインキュベーションが終了したら、細胞は、上述した濃度を用いて本発明の処方対象の有効成分と共に250μMのt−BOOHへ曝露した。24時間が経過したら、細胞を回収し、1200rpmで5分間遠心分離し;次にマロニルジアルデヒド(MDA)の産生を測定するために液体を回収した。単層を洗浄し、タンパク質及びグルタチオンレベル(GSH)を評価するために凍結させた。未処理細胞は、基礎酸化のコントロールとして使用し、t−ブチルヒドロペルオキシドで排他的に処理した細胞は、誘導酸化のコントロールとして使用した。
実施例4の抽出物及びコントロール物質(生理食塩水)は、以下の条件:
種:アルビノマウス(ハツカネズミ)
近交系:Balb/C/CNPB
動物の数:実験群当たり10匹の動物
性別:雄及び雌
体重:20〜25g
グループI(処理群):これらの動物には、2,000mg/kgの実施例4の抽出物の用量を投与
グループII(コントロール):これらの動物には、溶媒或いは生理食塩水(抽出物容積と同量)を投与
で、胃内カテーテルを用いて経口的に投与した。
実施例4の抽出物及びコントロール物質(生理食塩水)は、以下の条件:
種:アルビノラット(Rattus novergicus)
近交系:Holtzmann
動物の数:実験群当たり3匹の動物
性別:雄及び雌
体重:120〜160g
グループI(処理群):これらの動物には、2,000mg/kgの用量の実施例4の抽出物を投与
グループII(コントロール):これらの動物には、溶媒或いは生理食塩水(抽出物容積と同量)を投与
に従って、胃内カテーテルを用いて経口的に投与した。
Claims (22)
- 肝障害を治療するためのハーブ組成物であって、少なくとも1種のCordia(イヌヂシャ属)、Annona(バンレイシ属)或いはCurcuma(ウコン属)、若しくはそれらの組み合わせを有するものである、ハーブ組成物。
- 請求項1記載のハーブ組成物において、前記少なくとも1種は、イヌヂシャ属植物の1種である、ハーブ組成物。
- イヌヂシャ属、バンレイシ属及びウコン属植物のそれぞれの1種を含む、請求項1記載のハーブ組成物。
- イヌヂシャ属の花、バンレイシ属の葉、或いはウコン属の根、若しくはそれらの組み合わせを含む、請求項1記載のハーブ組成物。
- Cordia lutea(ムユヨ)の花、Annona muricata(トゲバンレイシ)の葉、及びCurcuma longa(ウコン)の根を含む、請求項1記載のハーブ組成物。
- 肝障害を治療するためのハーブ組成物であって、少なくとも1種のCordia(イヌヂシャ属)、Annona(バンレイシ属)或いはCurcuma(ウコン属)の抽出物、若しくはそれらの組み合わせを有するものである、ハーブ組成物。
- 請求項6記載のハーブ組成物において、前記少なくとも1種は、イヌヂシャ属植物の1種である、ハーブ組成物。
- イヌヂシャ属、バンレイシ属及びウコン属植物のそれぞれの1種の抽出物を含む、請求項6記載のハーブ組成物。
- Cordia lutea(ムユヨ)の花、Annona muricata(トゲバンレイシ)の葉、或いはCurcuma longa(ウコン)の根の抽出物、若しくはそれらの組み合わせを含む、請求項6記載のハーブ組成物。
- ムユヨの花、トゲバンレイシの葉、及びウコンの根の抽出物を含む、請求項6記載のハーブ組成物。
- 請求項10記載のハーブ組成物において、ムユヨの花、トゲバンレイシの葉、及びウコンの根の抽出物は、それぞれ約1:1:1〜約8:1:1の重量比で存在するものである、ハーブ組成物。
- 請求項10記載のハーブ組成物において、ムユヨの花、トゲバンレイシの葉、及びウコンの根の抽出物は、それぞれ約1:0.025〜1:0.025〜1の重量比で存在するものである、ハーブ組成物。
- 請求項10記載のハーブ組成物において、ムユヨの花、トゲバンレイシの葉、及びウコンの根の抽出物は、それぞれ約8:1:1の重量比で存在するものである、ハーブ組成物。
- 請求項13記載のハーブ組成物において、前記抽出物は含水アルコール抽出物である、ハーブ組成物。
- ハーブ組成物の調合に使用するための、選択された植物器官から含水アルコール抽出物を得るための方法であって、
前記選択された植物器官を乾燥させ、約0.35mm〜約0.1mmの範囲の粒子サイズを有する粉末を得るために前記乾燥させた植物器官を粉砕する工程と、
約6〜約8日間、ほぼ室温で含水アルコール溶液に前記粉末を漬け込む工程であって、これにより前記植物器官の抽出物を得るものである、前記漬け込む工程と、
前記抽出物を濃縮する工程と、
前記抽出物を凍結乾燥させ滅菌する工程と
を有するものである、方法。 - 請求項15記載の方法において、前記抽出物は、ロートエバポレータにおいて濃縮されるものである、方法。
- 患者における肝障害の治療或いは予防のための方法であって、治療上有効な量の請求項1〜14に記載されたハーブ組成物を患者に投与する工程を有するものである、方法。
- 請求項17記載の方法において、前記肝障害は、ウイルス感染によって引き起こされるものである、方法。
- 請求項18記載の方法において、前記ウイルス感染は、B型肝炎ウイルス、C型肝炎ウイルス、或いはその両者の組み合わせによって引き起こされるものである、方法。
- 請求項17記載の方法において、前記肝障害は、非ウイルス性肝炎である、方法。
- 請求項17記載の方法において、前記肝障害は、肝臓の線維化或いは肝硬変である、方法。
- フリーラジカルを中和し、患者におけるフリーラジカルの形成を予防するための方法であって、有効な量の請求項1〜14に記載されたハーブ組成物を、そのような治療が必要な患者に投与する工程を有するものである、方法。
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WO2004048358A1 (en) * | 2002-11-22 | 2004-06-10 | Nature's Sunshine Products, Inc. | Control of cancer with annonaceous extracts |
CN1593495A (zh) * | 2004-07-01 | 2005-03-16 | 邢乐道 | 一种治疗癌症的复方药物及其制备方法 |
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ES2154610B1 (es) * | 1999-09-23 | 2001-11-16 | Asac Compania De Biotecnologia | Nuevas actividades farmacologicas de los extractos de curcuma longa. |
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Cited By (2)
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JP2015143200A (ja) * | 2013-12-27 | 2015-08-06 | ハウスウェルネスフーズ株式会社 | Vcam−1発現抑制剤 |
KR101872622B1 (ko) * | 2016-12-20 | 2018-06-29 | 세종대학교산학협력단 | 니코틴 독성 해독용 그라비올라 추출물 제조방법 및 이 추출물을 함유하는 해독용 음료 조성물 |
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US20100260874A1 (en) | 2010-10-14 |
EP2205255B8 (en) | 2015-09-30 |
WO2009043176A9 (en) | 2009-07-02 |
BRPI0818178A2 (pt) | 2020-06-23 |
AR071731A1 (es) | 2010-07-14 |
MX2010003726A (es) | 2010-09-14 |
CA2701190C (en) | 2017-12-05 |
EP2205255B1 (en) | 2015-06-10 |
HK1145995A1 (en) | 2011-05-13 |
CA2701190A1 (en) | 2009-04-09 |
EP2205255A1 (en) | 2010-07-14 |
WO2009043176A1 (en) | 2009-04-09 |
US9775871B2 (en) | 2017-10-03 |
JP5486744B2 (ja) | 2014-05-07 |
BRPI0818178B1 (pt) | 2021-11-23 |
ES2546402T3 (es) | 2015-09-23 |
EP2205255A4 (en) | 2012-02-29 |
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